Ueno, Yuichiro (2009-05). THROUGH-BOND ENERGY TRANSFER CASSETTES FOR MULTIPLEXING & DEVELOPMENT OF METHODS FOR PROTEIN MONO-LABELING. Doctoral Dissertation. Thesis uri icon

abstract

  • A set of three through-bond energy transfer cassettes based on BODIPY as a donor and cyanine dyes as acceptors has been prepared via Sonogashira couplings, and their photophysical properties were examined. These cassettes fluoresce around 600 to 800 nm and are resolved by approximately 100 nm. This property is an important factor for multiplexing study in cellular imaging. Several useful fluorescent probes such as 5- and 6-carboxyfluorescein, water-soluble BODIPY, and water-soluble Nile Blue dyes, have also been synthesized and their photophysical properties studied. We have also attempted to develop a method for protein mono-labeling via a solidphase approach. The labeling of protein with one fluorescent dye facilitates quantification and single molecule imaging in biological applications. Various solidsupports such as PEGA, CPG, and BSA-coated CPG, were tested. Photolabile and chemically cleavable linkers were prepared to connect solid-supports and fluorophores. Unfortunately, our approach to the fluorescent mono-labeling of native proteins did not give us any conclusive results.
  • A set of three through-bond energy transfer cassettes based on BODIPY as a donor
    and cyanine dyes as acceptors has been prepared via Sonogashira couplings, and their
    photophysical properties were examined. These cassettes fluoresce around 600 to 800
    nm and are resolved by approximately 100 nm. This property is an important factor for
    multiplexing study in cellular imaging. Several useful fluorescent probes such as 5- and
    6-carboxyfluorescein, water-soluble BODIPY, and water-soluble Nile Blue dyes, have
    also been synthesized and their photophysical properties studied.
    We have also attempted to develop a method for protein mono-labeling via a solidphase
    approach. The labeling of protein with one fluorescent dye facilitates
    quantification and single molecule imaging in biological applications. Various solidsupports
    such as PEGA, CPG, and BSA-coated CPG, were tested. Photolabile and
    chemically cleavable linkers were prepared to connect solid-supports and fluorophores.
    Unfortunately, our approach to the fluorescent mono-labeling of native proteins did not
    give us any conclusive results.

publication date

  • May 2009